Table 2007.05C.
HPLC-UV parameters
| HPLC column | Zorbax SB-C18, 5 μm, 3.0 mm id × 25 cm, Agilent | ||
|---|---|---|---|
| Mobile phase A | 0.1% (v/v) trifluoroacetic acid-purified water | ||
| Mobile phase B | 0.1% (v/v) trifluoroacetic acid-acetonitrile | ||
| Pump program | |||
| Time, min |
%A |
%B |
Gradient curve |
| 0 | 80 | 20 | NAa |
| 25 | 30 | 70 | 1b |
| 30 | 0 | 100 | 1 |
| 31 | 80 | 20 | 1 |
| 40 |
80 |
20 |
NA |
| Detection | Ultraviolet absorbance detector (390 nm) | ||
| Column temperature, °C | 40 | ||
| Flow rate | Approximately 0.5 mL/minute; flow rate should be adjusted so that desired retention time will be obtained | ||
| Injected volume, μL | 25 | ||
| Injection time, min | 40 | ||
a
NA = Not applicable.
b
A gradient curve of 1 indicates a linear transition.